结核分枝杆菌furB基因的克隆、表达和纯化
结核分枝杆菌,,furB;表达;纯化;结核分枝杆菌,0引言,1材料和方法,2结果,3讨论,【参考文献】
Cloning, expression and purification of Mycobacterium tuberculosis furBJIANG Hong, XUE Ying, GAO Xue, SHI ChangHong, BAI YinLan, ZHANG Hai, LI Yuan, XU ZhiKai
Department of Microbiology, School of Basic Medicine, Fourth Military Medical University, Xian 710033, China
【Abstract】 AIM: To obtain Mycobacterium tuberculosis (MTB) furB from H37RvDNA, express efficiently in E. coli and purify the FurB proteins. METHODS: furB was amplified by PCR from the genome of H37Rv and cloned into pGEMTEasy vector. After sequenced, furB was recombinated into the expression vector pQE80L by restriction enzyme digestion, which was named pQE80LfurB. The plasmid pQE80LfurB was transformed into E.coli DH5α and induced by IPTG. The furB expression was analyzed by SDSPAGE and confirmed by Western blot with micespecific mAb against (His)6. RESULTS: furB was identical to what reported by Genbank. The pQE80LfurB vector expressed protein with relative molecule mass 15.0×103, which could be caught by (His)6 mAb. The expressed protein could be purified by NiNTA system kit with denatured condition. CONCLUSION: furB has been successfully cloned and efficiently expressed in E.coli, The results lay the basis for further study of FurB functions. ......
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